Renewal of embryonic and neonatal-derived cardiac-resident macrophages in response to environmental cues abrogated their potential to promote cardiomyocyte proliferation via Jagged-1-Notch1

Cardiac-resident macrophages (CRMs) play important roles in homeostasis, cardiac function, and remodeling. Although CRMs play critical roles in cardiac regeneration of neonatal mice, their roles are yet to be fully elucidated. Therefore, this study aimed to investigate the dynamic changes of CRMs during cardiac ontogeny and analyze the phenotypic and functional properties of CRMs in the promotion of cardiac regeneration. During mouse cardiac ontogeny, four CRM subsets exist successively: CX3CR1+CCR2-Ly6C-MHCII- (MP1), CX3CR1lowCCR2lowLy6C-MHCII- (MP2), CX3CR1-CCR2+Ly6C+MHCII- (MP3), and CX3CR1+CCR2-Ly6C-MHCII+ (MP4). MP1 cluster has different derivations (yolk sac, fetal liver, and bone marrow) and multiple functions population. Embryonic and neonatal-derived-MP1 directly promoted cardiomyocyte proliferation through Jagged-1-Notch1 axis and significantly ameliorated cardiac injury following myocardial infarction. MP2/3 subsets could survive throughout adulthood. MP4, the main population in adult mouse hearts, contributed to inflammation. During ontogeny, MP1 can convert into MP4 triggered by changes in the cellular redox state. These findings delineate the evolutionary dynamics of CRMs under physiological conditions and found direct evidence that embryonic and neonatal-derived CRMs regulate cardiomyocyte proliferation. Our findings also shed light on cardiac repair following injury.

IL-10 producing regulatory B cells ameliorated cardiac hypertrophy of viral myocarditis mice / 中华微生物学和免疫学杂志

Objective:To explore the mechanism of B10 cell involved in cardiomyocyte hypertrophy following myocarditis, and to develop potential therapeutic strategies.Methods:BALB/c mice infected with Coxsackie virus B3 induced viral myocarditis model. The expression of angiotensin (ANG)Ⅱ and its receptor in myocarditis mice was detected. The changes of B10 cells in the hearts of control mice and myocarditis mice were analyzed by flow cytometry. After losartan was administered to myocarditis mice, the degree of myocardial inflammation was detected by HE staining, the expression of inflammatory factors was detected by ELISA, the myocardial hypertrophy was detected by wheat germ agglutinin (WGA) staining, and the changes of B10 cells in the heart were analyzed by flow cytometry. The levels of cardiac troponin T (C-TNT) and high mobility group box 1 (HMGB1) protein in neonatal mouse cardiomyocytes treated with ANGⅡ and ANGⅡ+ IL-10 were detected. Cardiomyocytes were treated with ANGⅡ, ANGⅡ+ B10 cells, ANGⅡ+ B10 cells + IL-10 receptor antibody and ANGⅡ+ B cells to detect C-TNT protein levels, and Annexin-V/PI was used to detect the apoptosis of cardiomyocytes. Cardiomyocytes were treated with oxidized HMGB1, reduced HMGB1 and disulfide HMGB1, and C-TNT expression was detected.Results:Coxsackievirus B3 infection caused cardiac hypertrophy, high expression of ANGⅡ and its receptor, and transient increase of B10 cells in mice. Losartan treatment blocked the angiotensin receptor, reduced expansion of B10 cells. B10 cells alleviated apoptosis of cardiomyocytes and inhibited the production of HMGB1 induced by ANGⅡ patch by producing IL-10, thus alleviating viral myocarditis and cardiac hypertrophy.Conclusions:B10 cells may play an important role in myocardial protection in myocarditis.

Correlations of plasma eotaxin-2, sTNFR Ⅱ and other cytokines levels with the status of metabolic syndrome in type 2 diabetic nephropathy patients treated with maintenance hemodialysis / 临床检验杂志

Objective To investigate the correlations of plasma eotaxin-2,soluble tumor necrosis factor receptor(sTNFR) Ⅱ and other cytokines levels with the status of metabolic syndrome(MS) in type 2 diabetic nephropathy (DN) patients treated with maintenance hemodialysis(MHD).Methods Thirty type 2 DN patients with stable pathogenetic conditions and MHD treatment for more than 3 months,thirty newly diagnosed and untreated type 2 diabetes(T2D) patients and thirty healthy volunteers were enrolled in the study.The MS related markers,including blood pressure,waist circumference,body mass index,triglyceride(TG),high density lipoprotein cholesterol(HDL-C),fasting blood glucose (FBG),total cholesterol(TC),low density lipoprotein cholesterol(LDL-C) and C reactive protein(CRP),were determined.The expression levels of 40 cytokines were detected with the AAH-INF-G3 antibody microarray,and their correlations with MS related markers were analyzed by the Pearson method.Results The incidence rates of MS in DN patients with MHD and T2D patients were 88.89％ and 33.33％,respectively.Compared with T2D patients and healthy controls,the DN patients had more MS related markers,higher waist circumference and lower body mass index,and their plasma eotaxin-2,I-309 and sTNFR Ⅰ / Ⅱ levels increased significantly.Pearson correlation analysis showed that plasma eotaxin-2 and I-309 levels were positively related to MS risk factors such as TG,FBG and diastolic blood pressure(DBP) levels.Plasma sTNFR Ⅱ and I-309 levels were significantly positively correlated with plasma CRP levels.Conclusion A micro inflammatory state exists in type 2 DN patients with MHD.Abnormal glycolipid metabolism may influence on the immunologic and physiological state of human body,and then form the micro inflammatory state.Eotaxin-2 and I-309 may participate in this chronic and persistent process and further induce renal damage by upregulating sTNFR Ⅰ / Ⅱ levels,which may result in the formation of MS state in type 2 DN patients with MHD.

Intracellular trafficking of TREM2 is regulated by presenilin 1

Genetic mutations in triggering receptor expressed on myeloid cells 2 (TREM2) have been linked to a variety of neurodegenerative diseases including Alzheimer’s disease, amyotrophic lateral sclerosis, frontotemporal dementia and Parkinson’s disease. In the brain, TREM2 is highly expressed on the cell surface of microglia, where it can transduce signals to regulate microglial functions such as phagocytosis. To date, mechanisms underlying intracellular trafficking of TREM2 remain elusive. Mutations in the presenilin 1 (PS1) catalytic subunit of the γ-secretase complex have been associated with increased generation of the amyloidogenic Aβ (amyloid-β) 42 peptide through cleavage of the Aβ precursor amyloid precursor protein. Here we found that TREM2 interacts with PS1 in a manner independent of γ-secretase activity. Mutations in TREM2 alter its subcellular localization and affects its interaction with PS1. Upregulation of PS1 reduces, whereas downregulation of PS1 increases, steady-state levels of cell surface TREM2. Furthermore, PS1 overexpression results in attenuated phagocytic uptake of Aβ by microglia, which is reversed by TREM2 overexpression. Our data indicate a novel role for PS1 in regulating TREM2 intracellular trafficking and pathophysiological function.

Role of interleukin-17 in spinal dorsal horns in neuropathic pain in rats and its effect on activation of astrocytes / 中华麻醉学杂志

Objective To investigate the role of interleukin-17 (IL-17) in spinal dorsal horns in neuropathic pain (NP) in rats and its effect on activation of astrocytes.Methods In vivo experiment Sixty-four male SPF Sprague-Dawley rats,aged 6-8 weeks,weighing 180-200 g,were randomly divided into 3 groups using a random number table:control group (group C,n =16),sham operation group (group S,n =24) and group NP (n =24).The animals were anesthetized with intraperitoneal pentobarbital sodium,the L5,6 spinal nerves of the left side of the rat were gently separated and exposed,tightly ligated with 5-0 silk suture and transected.In group S,the L5,6 spinal nerves of the left side of the rat were only exposed.In group C,no operation was performed.Mechanical pain threshold was measured at day 1 before operation and days 1,3,5,7,10 and 14 after operation.The expression of IL-17,IL-6,IL-1β and tumor necrosis factor-alpha (TNF-α) mRNA in the spinal dorsal horn was determined using quantitative real-time PCR at day 7 and day 14 after operation.At day 7 after operation,the activation of astrocytes in the spinal dorsal horn was detected.In vitro experiment Primarily cultured astrocytes of neonatal rats were randomly divided into 4 groups using a random number table:control group (group C,n=22),10 ng/ml IL-17 group (I10 group,n=18),50 ng/ml IL-17 group (I50 group,n-18) and 100 ng/ml IL-17 group (I100 group,n=22).In I10,I50 and I100 groups,the astrocytes were incubated with the culture medium containing 10,50 and 100 ng/ml IL-17,respectively.The proliferation of astrocytes was detected by MTT at 24,48 and 72 h of incutation or culture.The expression of IL-6,IL-1β and TNF-α mRNA was determined using quantitative real-time PCR.Results In vivo experiment Compared with group C,the mechanical pain threshold was significantly decreased at 3-14 days after operation,the expression of IL-17,IL-6 and IL-1β mRNA in the spinal dorsal horn was up-regualted at 7 days after operation,and the activation of astrocytes was increased in group NP,and no significant change was detected in the mechanical pain threshold at each time point after operation in group S.In vitro experiment Compared with group C,the proliferation of astrocytes was significantly increased at 48 h of incubation in I10 and I50 groups,the proliferation of astrocytes was significantly increased at 48 and 72 h of incubation,and the expression of IL-6 and IL-1β mRNA was up-regulated in I100 group,and no significant change was found in the proliferation of astrocytes in group S.Conclusion Up-regulated expression of IL-17 in spinal dorsal horns may be involved in the maintenance of NP,and the mechanism is related to promoted activation of astrocytes and induced inflammatory responses in rats.

Epidemiological study of the genes conferring aminoglycoside resistance in 36 strains of multidrug-resistant Acinetobacter baumannii / 中国感染与化疗杂志

Objective To investigate the prevalence of genes conferring aminoglycoside resistance in multidrug-resistant strains of Acinetobacter baumannii (MDR-ABA).Methods Multidrug-resistant A.baumannii strains were isolated during the period from August to November 2012 from patients in the affiliated hospital of Jiangsu University and the First Hospital of Zhen-jiang.Kirby-Bauer diffusion method was used to determine the susceptibility of these strains to antimicrobial agents.PCR was performed to detect the aminoglycoside resistance genes.Results The 36 MDR-ABA strains showed high resistance rates to most antimicrobial agents except cefoperazone-sulbactam.The prevalence of the genes conferring aminoglycoside resistance, aac (3)-I,aac (6’)-Ib,aph (3’)-I and armA,was 72.2% (26/36),72.2% (26/36),80.6% (29/36)and 80.6% (29/36), respectively.Conclusions The MDR-ABA strains in this study are highly resistant to antimicrobial agents,which is closely as-sociated with the genes conferring aminoglycoside resistance.

Increased frequency of Th17 cell in patients with Hashimoto's thyroiditis / 中华内分泌代谢杂志

Objective To study the alteration of Th17 cells and related molecules in patients with Hashimoto's thyroiditis (HT).Methods Th17 cells were determined by flow cytometry.Real-time PCR method was applied to detect the expression of the orphan nuclear receptor RORγt and interleukin(IL) -17.Serum IL-6 and IL-23 were detected by ELISA method.Results ( 1 ) Compared with healthy controls,the frequency of Th17 cells [ (0.75± 0.79) ％ vs ( 0.28 ± 0.23 )％ ] and expression of RORγt ( 0.30 ± 0.38 vs 0.04 ± 0.02,both P ＜ 0.05 ) were significantly increased in HT patients.( 2 ) The levels of IL-6 and IL23 in HT patients were higher than those in healthy controls ( 3.66 ± 4.70 vs 0.47 ± 1.11,154.7 ± 75.81 vs 80.65 ± 61.41,both P＜0.05 ).( 3 ) A positive correlation between Th17 cells and serum TgAb was revealed in HT patients ( r =0.848 4,P =0.007 7 ).(4) The results of PCR showed that IL-17 and RORγt expressed in thyroid tissues of patients HT.Conclusion An increased frequency of Thl7 cells was found in HT patients,implying that this cell subset may play an important role in the pathogenesis of autoimmune thyroid disease.

IL-4 can inhibit il-17 production in collagen induced arthritis

IL-4 is a cytokine that induces differentiation of naive helper T cells into Th2 cells. Once activated by IL-4, Th2 cells subsequently produce additional IL-4. To examine the effect of IL-4 on IL-17 production and its effect in Collagen-Induced Arthritis [CIA] mice. Method: In this study, a chicken collagen-II-induced experimental arthritis [CIA] model was used in DBA/1 mice to investigate the relationship between IL-4 and IL-17 as well as other inflammatory factors. On the 38th day after the mice were induced with CIA, the expression of IL-17 and IL-4 as well as IFN-gamma and IL-13 in sera of the mice was measured by QRT-PCR and ELISA. The result of QRT-PCR analysis of IL-17 and IL-4 mRNA levels in the spleen showed that IL-17 is increased significantly at the onset of CIA in the spleen [p<0.01]. Meanwhile, IL-17 is generally reduced at the peak of CIA but IL-4 is increased significantly at this peak in the spleen [p<0.05] when the weight of the animal was taken into consideration. IL-4 can be involved in the production of IL-17 at especially the peak of CIA. These results imply that the inhibition of IL-17 may decrease the expression of IL-lp and IL-6 production which will result in the aggravation of arthritis

The detection and significance of high mobility group box chromosomal protein 1, RORγt and inter leukin-17 in peripheral blood of rheumatoid arthritis / 中华风湿病学杂志

Objective To detect the expression levels of high mobility group box chromosomal protein 1 (HMGB1) and Th17 cells transcription factors, related cytokines in peripheral blood of rheumatoid arthritis (RA) patients and analyze the relations between HMGB1 and CRP, ESR, RF in RA patients. The other aim of this study is to identify the expression level of HMGBI and the relationship between HMGB1 and Th17 in RA patients. Methods The mRNA levels of HMGB1, RORyt, interleukin (IL)-17 in the peripheral blood mononuclear cells (PBMC) were determined by quantitative real-time PCR (QRT-PCR) from 80 patients with rheumatoid arthritis,including 32 RA patients in stable phase and 48 patients in active phase, and 50 healthy volunteers. The concentration of HMGB1, IL-23, IL-17 in plasma were detected by enzyme linked immunosorbent assay (ELISA), one-way ANOVA and Spearman's correleation were adopted for statistical analysis.Results The mRNAs of HMGBI, RORyt and IL-17 in RA patients were higher than that in healthy control group (P<0.05), especially in active RA patients [ HMGB 1 (0.424±0.262) pg/ml, RORγt (0.34±0.25) pg/ml,IL-17 (1.42±0.38) pg/ml,P<0.01 ] when compared with patients with stable disease. The concentration of HMGB1, IL-23 and IL-17 in the plasma of RA patients was higher than that of the healthy control group (P< 0.05), and was positively correlated with the expression levels of HMGB1, Th 17-associated factors and the level of CRP, ESR, RF in RA patients' plasma(P<0.05). Conclusion The HMGB1 and Thl7 cells levels are higher in active RA patients than those in patients with stable disease, arid there is significant positive correlation between them. Detection of peripheral HMGB1 and Thl7 cell-specific transcription factors or related cytokines can help to understand the development and progress of rheumatoid arthritis and provide clues for new treatment targets for RA.

Adeno-associated virus mediated T-bet gene transfer into SGC-7901 cell to regulate IFN-gamma production / 生物医学工程学杂志

In order to investigate the effect of T-bet on malignant cells, we selected SGC-7901, a kind of human gastric carcinoma cell line, and used gene clone technique and adeno-associated virus (AAV) packing technology, thus obtaining a recombinant rAAV-eGFP-T-bet and T-bet gene-transfected SGC-7901 cells. Then the function of T-bet gene-infected SGC-7901 cells was researched by detecting the levels of IFN-gamma and T-bet production. The results showed: (1) It was verified that rAAV-T-bet's packing was completed; (2) After SGC-7901 cells was transfected by rAAV-eGFP-T-bet, a green fluorescence was found in about 30%-40% SGC-7901s, and the gene of 1670 bp (T-bet) and 388 bp (IFN-gamma) were generated from SGC-7901s cells; (3) The proteins of IFN-gamma and T-bet secreted by SGC-7901 cells were also detected. These reveal that SGC-7901 cell is efficiently infected by rAAV encoding T-bet, which can induce transfected cells to secret IFN-gamma. It may be useful in the researches on cancer immune therapy of transfecting T-bet gene.

Study on the interaction between presenilin 1 and carboxyl terminus of Hsc70 interacting protein / 中华老年医学杂志

ObjectiveTo understand the pathological and physiological roles of Presenilin 1 (PS1) in Alzheimer's disease (AD) recurrence, and the interaction between PSI and carhoxyl terminus of Hsc70 interacting protein (CHIP). MethodsThe yeast two-hybrid system was applied to identify a novel PS1 interacting protein as CHIP. After pGBKT7-PS1-C203 bait plasmid and full fragement CHIP of pACT2-CHIP expression vector were constructed, the interaction between PSI and CHIP was tested by β-galactosidase assay, pGBKT7-PS1-C203 was co-transfected with pACT2-CHIP into 293T cells and the interaction between PS1 and CHIP was tested by co-immunoprecipitation and Western blot. ResultsSpecificity of the interaction between PS1 and CHIP was identified by β-galactosidase assay and co- immunoprecipitation. ConclusionsCHIP is able to modulate chaperone functions and the pathway of protein ubiquitination/degradation. CHIP may regulate a proper assembly of the γ-secretase complex through its interaction with PSI, which is helpful to elucidate the mechanism of AD pathology.

The distribution and structure of class Ⅰ integron in the multidrug-reisistant Pseudomonas aeruginosa / 中华检验医学杂志

Objective To investigate the antimicrobial susceptibility of Pseudomonas aeruginosa (P. aeruginosa) isolated from Zhenjiang area to 13 routinely used antibiotics and identify the structure and dissemination of class Ⅰ integron. Methods K-B test was used to determine the resistant rate of 71 strains of P. aeruginosa. DNA template was extracted by boiling method, PCR method was utilized to detect class Ⅰintegron, and subsequently gene cassettes were analyzed by sequencing. Results The resistant rates to 13 routinely used antibiotics were quite different from 18. 3 to 77.5% among 71 strains of P. aeruginosa. The prevalence of class Ⅰ integron was 38%. These integrons include 5 gene cassettes ( aadB, aac (6) - Ⅱ , PSE-Ⅰ , dfrA17 and aadAS), in which dfrA17 and aadA5 gene cassette were frequently found. Comparing with the negative strains of integron, the positive strains of integron has obviously higher resistance to ten the antibiotics including piporacillin, piperacillin-tazobactam, ceftriaxone, cefepime, ceftazidime, gentamicin,amikacin, tobmmycin, levofloxacin, and ciprofloxacin. Conclusions The resistant rates of P. aeruginosa to 13 drugs were different, and the resistant rates of integron positive strains were obviously higher than integron negative strains, which indicates that integron may play an important role in multidrug reisistance of P. aeruginoosa.

Avian Influenza: Should China Be Alarmed?

Avian influenza has emerged as one of the primary public health concern of the 21st century. Influenza strain H5N1 is capable of incidentally infecting humans and other mammals. Since their reemergence in 2003, highly pathogenic avian influenza A (H5N1) viruses have been transmitted from poultry to humans (by direct or indirect contact with infected birds) in several provinces of Mainland China, which has resulted in 22 cases of human infection and has created repercussions for the Chinese economy. People have been concerned whether a new pandemic will occur in the future. The eradication of pathogenic avian influenza viruses appears to be the most effective way to prevent an influenza pandemic. This paper will examine the features of H5N1, including incidence, infection, immunity, clinical management, prevention and control, and therapy in Mainland China.

Study on Optimizing Preparative Procedure for Isolating and Purifying the Soluble Collagen Type Ⅱ / 临床检验杂志

Objectives To establish a reliable method for isolating and purifying soluble collagen type Ⅱ(SCⅡ)by optimizing preparative procedure.Method The chicken sternal cartilage was selected as raw material.Guanidine hydrochloride was used to remove the proteoglycans.The digestion manners of pepsin,sodium chloride concentrations for salting,types of DEAE anion resin were studied for extracting SCⅡ.The SCⅡ identification was made by SDS-PAGE,absorption spectrum and amino acid analysis.Result It was convenient for pre-treatments of chicken sternal cartilage.The proteoglycans could be efficiently removed by 4 mol/L guanidine hydrochloride.The satisfied results were obtained by limited enzyme digestion of pepsin added by two steps.The optimizing concentration of sodium chloride for salting was 2.4mol/L.SDS-PAGE maps revealed that the bands of purified SCⅡand standard CⅡ were at the same location.The absorption peak of SCⅡ was at 230nm.The concentrations of Gly,Pro and Ala were the highest in 15 amino acids.Conclusion The improved method has significant advantages of simple working process,result reliability and convenient source of raw material.It is suitable for purifying the SCⅡ at variable scales in research works and clinic application.The SCⅡ product obtained has high purity and accords with the characteristics of collagen type Ⅱ.

Studies on B cell differentiation from cord blood CD34~+CD19~- hematopoietic stem/progenitor cells in vitro / 中国免疫学杂志

Objective:To investigate the condition of B cell differentiation from cord blood CD34+CD19-hematopoietic stem/progenitor cells in vitro.Methods:CD34+CD19-cells from cord blood were isolated and purified by using immunomagnetic beads separation system.CD34+CD19-cells supported by murine S-17 stromal cells were stimulated in co-culture with T3 and cytokines.Differentiated B cells were analyzed by flow cytometry.Results:The amplification of the B cells derived from CD34+CD19-hematopoietic stem/progenitor cells in co-culture with T3 and IL-7 reached 198-fold of increase,most of the induced cells expressed CD10 and CD19.Conclusion:In the experimental conditions selected,co-culture of CD34+CD19-cells with T3,IL-7 and murine S-17 stromal cells could stimulate differentiate toward to B cells in vitro.

The relation of hypoxia-inducible factor with inflammation and tumor / 中国药理学通报

Regional microenviroment hypoxia is a common feature in inflammation and malignancy. Cells are able to trigger an adaptive response to hypoxia conditions that result in hypoxia-inducible factor(HIF)overexpression. Based on a brief description of the structure and functions of HIF, this article discusses the relationship between inflammation, tumor and HIF.